TGI-RTase has reverse transcriptase activity that depends on DNA/RNA hybrid chain and can rapidly synthesize cDNA. This enzyme has strong strand switching activity. Therefore, the enzyme can be used for synchronous reverse transcription and ligation of nucleic acid molecules such as tRNA, ncRNA, and miRNA (TGIRT template-switching reaction, for TGIRT-Seq).
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: TGIRT-1 (for 1nmol)
Cat. No.: TGIRT-10 (for 10nmol)
TGI-RTase has reverse transcriptase activity that depends on DNA/RNA hybrid chain and can rapidly synthesize cDNA. This enzyme has strong strand switching activity. Therefore, the enzyme can be used for synchronous reverse transcription and ligation of nucleic acid molecules such as tRNA, ncRNA, and miRNA (TGIRT template-switching reaction, for TGIRT-Seq). TGI-RTase has strong extensibility, so it is easier to obtain full-length cDNA. In addition, the strong switch activity of this enzyme makes it possible to use the 5' end cap structure of RNA for SMART Library Building (SMART-Seq). The reaction temperature of the enzyme is 37-60°C, and the optimal reaction temperature is 50°C.
TGI-RTnase (10 pmol/μl): 100 μl
5×TGI-RTase Buffer: 1 ml
100mM DTT: 0.5 ml
Higher thermal stability, persistence, and fidelity than retroviral reverse transcriptase allow full-length end-to-end cDNA synthesis from highly structured or strictly modified RNA.
New end-to-end template-switching activity, which can connect RNA-seq or PCR linkers during reverse transcription, and no longer requires a separate RNA 3'- linker connecting step
No RNA ligase is required, and the requirement for template quantity is low, which simplifies the steps of RNA-seq library construction and makes it more efficient.
Strand-specific transcriptome analysis
Whole-cell or cell-free transcriptome sequencing
Noncoding RNA transcriptome sequencing
RIP-seq, HITS-CLIP, irCLIP, and CRAC
Identification of RNA base modifications by high throughput sequencing
RNA structure mapping
Synthesis of long cDNA
Genome-wide or targeted RNA structure analysis
The minimum shelf life is 2 years at -20°C and 3 months at 4°C.
5×TGI-RTase Buffer 4 μl 100mM DTT 2 μl RNA 1~50ng 1μM of Annealed DNA/RNA duplex 2 μl TGI-RTnase (10 pmol/μl) 1 μl RNase Free H2O up to 17.5 μl
After 30 min incubation at room temperature, add 2.5 μl of 10mM dNTP and react for 5-60 min (usually 15 min) at 37-60°C (the recommended temperature is 50 ° C).
After the reaction, add 1 μl 5N NaOH and heat at 95°C for 3 min to inactivate the enzyme. After inactivation, add 5N HCl to neutralize the high alkaline conditions.
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